Interpretive Summary: Conservation of the biodiversity of fruit crops is important to the future of horticulture in Kazakhstan. A field collection of fruit germplasm with more than 4000 cultivars and wild selections is grown in the Pomological Garden of the Institute of Horticulture and Viticulture near Almaty, to preserve the native species, as well as cultivated fruit. These plants are at risk from environmental as well as biotic threats. Cryopreservation in liquid nitrogen (-196°C) is used for reliable long-term preservation of plant genetic resources. A cryopreserved collection allows preservation for an unlimited term, retention of viability, regeneration potential and genetic stability and provides a backup of the collection that is secure from human or environmental factors, or attack by insects and diseases. A cryo genebank is now being established based on experiments that defined optimal cryopreservation approaches for liquid nitrogen (-196°') storage of these cultures.The cryogenic collection includes the following plants stored as dormant buds: 124 apple genotypes with recovery from 92% to 100%; 60 sweet cherry genotypeswith recovery from 62% to 75%; 17 pear genotypeswith recovery from 90% to 100%; and 25 sour cherry genotypes with recovery from 75% to 82%. Collections stored as in vitro shoot tips include: 5 pear genotypes with 83.3% to 84.6 % regrowth; 6 cherry genotypes with 40% to 80% regrowth; 15 black currant genotypes with 46.7% to 100% regrowth; 25 raspberry genotypes with 50% to 80% regrowth; and 28 strawberry genotypes with 40% to 75% regrowth. This collection is the beginning of a national germplasm storage system for Kazakhstan.

Technical Abstract:
Conservation of the biodiversity of fruit crops is important to the future of horticulture in Kazakhstan. A field collection of fruit germplasm with more than 4000 cultivars and wild selections is grown in the Pomological Garden of the Institute of Horticulture and Viticulture near Almaty, to preserve the native species, as well as cultivated fruit. These plants are at risk from environmental as well as biotic threats. Cryopreservation in liquid nitrogen (-196°C) is used for reliable long-term preservation of plant genetic resources. A cryopreserved collection allows preservation for an unlimited term, retention of viability, regeneration potential and genetic stability and provides a backup of the collection that is secure from anthropogenic and abiotic factors, or attack by insects and diseases. A cryo genebank is now being established based on experiments that defined optimal cryopreservation approaches for liquid nitrogen (-196°') storage of these cultures. A PVS2 vitrification protocol was used with in vitro cultured shoot tips. Shoot tips (~ 1 mm) were precultured for 2 days in cold acclimation conditions on MS medium with 0.3 ' sucrose. Shoot tips were then placed in loading solution of 2 ' glycerol in 0.4 ' sucrose for 20 min, then placed in 1.2 ml cryovials on ice with 1 ml PVS2 for 80 min and submerged in liquid nitrogen. Rewarming of samples was carried out in a 45°C water bath for 1 min, then in 22°C water for 1 min. Dormant buds were cryopreserved with the following protocol: Sion wood was collected after the temperature -10 ' and below. The acclimated scions were dehydrated to 30% moisture at -3 oC to -5 oC, then cooled at 1°'/hour to –25°' and held at this temperature for 24 hours before placing in vapor phase over LN at -150°'. Scions were rewarmed for 24 hr in cold room at +4oC. Buds from the scions were grafted onto rootstocks or regenerated in vitro. The cryogenic collection includes the following plants stored as dormant buds: 124 apple genotypes [Malus x domestica Borkh. and Malus sieversii (Ledeb.) M. Roem)] (cultivars, hybrids, wild forms) with recovery from 92% to 100%; 60 sweet cherry genotypes (Prunus avium L.) with recovery from 62% to 75%; 17pear genotypes (Pyrus communis L.) with recovery from 90% to 100%; 25 cherry genotypes (Prunus cerasus L.) with recovery from 75% to 82%. Collections stored as shoot tips by PVS2 vitrification of in vitro shoot tips include: 5 pear genotypes (Pyrus communis L.) with 83.3% to 84.6 % regrowth; 6 cherry genotypes (Prunus cerasus L.), with 40% to 80% regrowth; 15 black currant genotypes (Ribes nigrum L.) with 46.7% to 100% regrowth; 25 raspberry genotypes (Rubus idaeus L., Rubus slesvicensis Lange) with 50% to 80% regrowth. Encapsulation-dehydration was used to cryopreserve 28 strawberry genotypes (Fragaria x ananassa Duch.) with 40% to 75% regrowth. This collection is the beginning of a national germplasm storage system for Kazakhstan.